PROJECT SUMMARY Acute survival following myocardial infarction has precipitated a growing heart failure epidemic, warranting de- velopment of disease-altering treatments to restore tissue integrity and ventricular function. The homeostatic capability for self-renewal in the adult human heart is insufficient in the setting of myocardial injury. Adult stem cell therapies, conceived to bolster this innate regenerative reserve and achieve clinically meaningful cardiac regeneration, have been found safe but demonstrate inconsistent benefit in ischemic cardiomyopathy. This recognized roadblock impedes the roll-out of cardiac regenerative medicine in clinical practice. To prime re- generative proficiency, our team initially introduced the lineage-specifying cardiopoiesis platform. Despite this optimization, a range of benefit following cardiopoietic stem cell therapy was experienced in two separate clini- cal trials. Exploiting the C-CURE and CHART-1 trial experience, preliminary work documented an altered mi- croRNA and secretome landscape germane to cardiopoietic stem cells that yielded the best outcomes in hu- man testing. Use of clinical readouts in conjunction with systems biology deconvolution of trial-biobanked car- diopoietic stem cell samples underpins an innovative approach to target pathways that drive regenerative ac- tion. The overarching hypothesis of the present proposal is that the molecular roadmap segregating high re- generative properties can be actuated to reverse engineer stem cells featuring enhanced regenerative profi- ciency. Thus, a clinical trial-informed paradigm is here introduced to dial-up the regenerative aptitude of stem cells. The proposal aims to: (1) engineer a highly regenerative cardiopoietic (hrCP) phenotype through targeted microRNA programming to achieve enhanced regenerative efficacy tested in murine ischemic cardiomyopathy; (2) establish mechanisms underlying hrCP action through a proteomic systems approach resolving impact on the disease substrate with focus on neovasculogenesis as a prioritized tissue compartment in repair; and final- ly (3) scale-up hrCP for translation in a porcine model of ischemic heart failure with molecular and functional imaging tracking biodistribution and therapeutic effectiveness, along with histological and high-throughput analysis, to demonstrate bioequivalency in clinically relevant conditions. Timely execution of stated aims is fa- cilitated by a multidisciplinary team with a track record in translating stem cell innovation, established method- ologies and resources, and a body of supporting preliminary work. On completion, the proposed research will realize the prospect of a clinomics-informed evolution in stem cell therapy for ischemic heart failure delivering an engineered and validated highly regenerative biotherapy.